Abstract
Long-term cultivation of human cells, including stem cells, can lead to essential transformations of the karyotype and genetic instability. The aim of this research was a comparative cytogenetic study of the karyotype of new human stem cell line 4BL at 160 and 205 passages. During a standard cytogenetic examination, the nullisomy and monosomy of chromosomes 10 and 13, monosomy of chromosomes 4, 8, 11, 15, 17, 21, and X; and t(1, 11), t (5, 15), t(12, 15), and t(16, 21) were observed; also, six regular marker chromosomes were detected. At 160 and 205 passages, the modal class of the karyotype was 42–43 chromosomes. While passaging increased frequency of polyploidy cells (from 2.8 to 36%), disappearance of nearhaploid cells (22.1% at the 160th passage) and a decreased level of early division of chromatids (from 5 to 1.5%) were observed. We assume the stabilization of the karyotype of cell line 4BL at 205 passages and consider that it is necessary to conduct an additional molecular and cytogenetic study for the objective identification of the number of chromosomes of the modal class, as well as the number of chromosomal anomalies, and for forecasting the direction of the karyotype evolution of human cells 4BL in vitro.
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Gaebel, R., Furlani, D., Sorg, H., et al., Cell origin of human mesenchymal stem cells determines a different healing performance in cardiac regeneration, PLoS One, 2011, vol. 6, no. 2, p. el5652.
Wagner, W., Bork, S., Lepperdinger, G., et al., How to track cellular aging of mesenchymal stromal cells? Aging (Albany, New York), 2010, vol. 2, no. 4, pp. 224–230.
Peters, R., Wolf, M.J., Broek, M., et al., Efficient generation of multipotent mesenchymal stem cells from umbilical cord blood in stroma-free liquid culture, PLoS One, 2010, vol. 5, no. 12, p. e15689.
Wagner, W., Ho, A.D., and Zenke, M., Different facets of aging in human mesenchymal stem cells, Tissue Engin., 2010, vol. 16, no. 4, pp. 445–453.
Furlani, D., Li, W., Pittermann, E., Klopsch, C., et al., A transformed cell population derived from cultured mesenchymal stem cells has no functional effect after transplantation into the injured heart, Cell Transplant., 2009, vol. 18, no. 3, pp. 319–331.
Kim, J., Kang, J.W., Park, J.H., et al., Biological characterization of long-term cultured human mesenchymal stem cells, Arch. Pharm. Res., 2009, vol. 32, no. 1, pp. 117–126.
Bernardo, M.E., Zaffaroni, N., Novara, F., et al., Human bone marrow derived mesenchymal stem cells do not undergo transformation after long-term in vitro culture and do not exhibit telomere maintenance mechanisms, Cancer Res., 2007, vol. 67, no. 19, pp. 9142–9149.
Maitra, A., Arking, D.E., Shivapurkar, N., et al., Genomic alterations in cultured human embryonic stem cells, Nat. Genet., 2005, vol. 37, no. 10, pp. 1099–1103.
Longo, L., Bygrave, A., Grosvels, F.G., and Pandolfi, P.P., The chromosome make up of mouse embryonic stem cell is predictive of somatic and germ cell chimerism, Transgenic Res., 1997, vol. 6, pp. 321–328.
Qin, Y., Ji, H., Wu, Y., and Liu, H., Chromosomal instability of murine adipose tissue-derived mesenchymal stem cells in long-term culture and development of cloned embryos, Cloning Stem Cells, 2009, vol. 11, no. 3, pp. 445–452.
Iatsyshyna, A.P., Genetic Instability of Mammalian Cells in vitro, Visn. Ukr. Tvar. Genet. Selekts., 2010, vol. 8, no. 1, pp. 165–178.
Lukash, L.L., Regulation of genome variability of mammalian somatic cells by exogenous biological factors, Biopolym. Cell, 2004, vol. 20, nos. 1/2, pp. 93–105.
Lukash, L.L., Iatsyshyna, A.P., Kushniruk, V.O., and Pidpala, O.V., Reprogramming of somatic cells of adult humans, in Factors of Experimental Evolution of Organisms, Kyiv: Logos, 2011, pp. 493–498.
Freshni, R.Ya., Kul’tura zhivotnykh kletok: Prakt. rukovodstvo (Animal Cell Culture: A Practical Guide), Moscow: Binom, 2010.
Hungerford, D.A., Leucocytes culture from small inocula of whole blood and the preparation of metaphase chromosomes by treatment with hypotonic KCl, Stain. Technol., 1965, vol. 10, no. 6, pp. 333–338.
Seabright, M.A., A rapid banding technique for human chromosomes, Lancet, 1971, vol. 2, pp. 971–972.
Shaffer, L.G., Slovak, M.L., and Campbell, L.J., ISCN 2009: An International System for Human Cytogenetic Nomenclature, Basel: Karger, 2009.
Akopyan, H., Sirenko, A., Sedneva, I., et al., Cytogenetic assay in apoptosis investigations, Folia Histochem. Cytobiol., 2001, vol. 39,suppl. 2, pp. 158–160.
Yih, L.H. and Lee, T.C., Induction of C-anaphase and diplochromosome through dysregulation of spindle assembly checkpoint by sodium arsenite in human fibroblasts, Cancer Res., 2003, vol. 63, no. 20, pp. 6680–6688.
Shinawi, M. and Cheung, S.W., The array CGH and clinical applications, Drug Discov. Today, 2008, vol. 13, nos. 17/18, pp. 760–770.
Iatsyshyna, A.P., Pidpala, O.V., Kochubei, T.P., and Lukash, L.L., Cytogenetic analysis of G1 spontaneously immortalized mouse cells, Biopolym. Cell, 2006, vol. 22, no. 4, pp. 299–306.
Glazko, T.T., Iatsyshyna, A.P., Pidpala, O.V., and Lukash, L.L., Succession of cytogenetic characteristics in passages of G1 mouse embryonic germ cells, Klet. Transplantol. Tkan. Inzhener., 2007, vol. 2, no. 3, pp. 47–50.
Lee, H.O., Davidson, J.M., and Duronio, R.J., Endoreplication: polyploidy with purpose, Genes Dev., 2009, vol. 23, no. 21, pp. 2461–2477.
Clouston, H.J., Herbert, M., Fenwic, J., et al., Cytogenetic analysis of human blastocysts, Pren. Diagn., 2002, vol. 22, no. 12, pp. 1143–1152.
Viuff, D., Palsgaard, A., Rickords, L., et al., Bovine embryos contain a higher proportion of polyploidy cells in the trophectoderm than in the embryonic disc, Mol. Reprod. Dev., 2002, vol. 62, no. 4, pp. 483–488.
Zybina, T.G. and Zybina, E.V., Cell reproduction and genome multiplication in the proliferative and invasive trophoblast cell populations of mammalian placenta, Cell. Biol. Int., 2005, vol. 29, no. 12, pp. 1071–1083.
Walen, K.N., The origin of transformed cells studies of spontaneous and induced cell transformation in cell cultures from marsupials, a snail, and human amniocytes, Cancer Genet. Cytogenet., 2002, vol. 133, no. 1, pp. 45–54.
Lozins’ka, M.R., Gnateiko, O.Z., and Gavrilyuk, Yu.I., The level of spontaneous genomic mutations in somatic human embryonic cells, Tsitol. Genet., 1994, vol. 26, no. 3, pp. 70–84.
Guleyuk, N.L., Cytogenetic characteristic of persons with disturbed menstrual function, Tsitol. Genet., 1994, vol. 26, no. 3, pp. 75–79.
Guleyuk, N.L., Zastavna, D.V., Bezkorovaina, G.M., and Akopyan, G.R., Efficiency of prenatal diagnosis of chromosomal pathologies and cytogenetic characteristics of cultured amniocytes, Eksp. Klin. Fiziol. Biokhim., 2003, no. 3, pp. 7–14.
Akopyan, G.R., Sirenko, A.G., Gnateiko, O.Z., et al., C-anaphase as a cytogenetic marker of cell apoptosis in acute lymphoblast leucosis in children, Eksp. Onkol., 1999, vol. 21, no. 2, pp. 127–132.
Akopyan, G.R., Early chromosome disjunction as an informative marker of chromosomal instability in human cells, in Advances and Problems of Genetics, Breeding, and Biotechnology, Kyiv: Logos, 2007, vol. 1, pp. 390–394.
Iatsyshyna, A.P., Kvasha, S.M., Pidpala, O.V., et al., Genetic instability of G1 mouse embryonic germ cells and disturbances of mitosis checkpoint functions in p53, Biopolym. Cell, 2007, vol. 23, no. 4, pp. 338–346.
Morgunkova, A.A., Almazov, V.P., Strunina, S.M., et al., Dominant-negative inactivation of p53: the effect of the proportion between a trans-dominant inhibitor and its target, Mol. Biol. (Moscow), 2003, vol. 37, no. 1, pp. 102–109.
Diaz, A., Elvira, G., and Silva, A., P53 regulates the proliferation, differentiation and spontaneous transformation of mesenchymal stem cells, Exp. Cell Res., 2009, vol. 315, no. 20, pp. 3598–3610.
Zielke, N., Querings, S., Rottig, C., et al., The anaphase-promoting complex/cyclosome (APC/C) is required for replication control in endoreplication cycles, Genes Dev., 2008, vol. 22, no. 12, pp. 1690–1703.
Wang, Z., Inuzuka, H., Fukushima, H., et al., Emerging roles of the FBW7 tumour suppressor in stem cell differentiation, EMBO Rep., 2011, vol. 13, no. 1, pp. 36–43.
Chircop, M., Malladi, C.S., Lian, A.T., et al., Calcineurin activity is required for the completion of cytokinesis, Cell Mol. Life Sci., 2010, vol. 67, no. 21, pp. 3725–3737.
Chircop, M., Sarcevic, B., Larsen, M.R., et al., Phosphorylation of dynamin II at serine-764 is associated with cytokinesis, Biochim. Biophys. Acta, 2011, vol. 1813, no. 10, pp. 1689–1699.
Rosario, C.O., Ko, M.A., Haffani, Y.Z., et al., Plk4 is required for cytokinesis and maintenance of chromosomal stability, Proc. Natl. Acad. Sci. U.S.A., 2010, vol. 107, no. 15, pp. 6888–6893.
Boudolf, V., Lammens, T., Boruc, J., et al., CDKB1,1 forms a functional complex with CYCA2,3 to suppress endocycle onset, Plant Physiol., 2009, vol. 150, pp. 1482–1493.
Lordier, L., Chang, Y., Jalil, A., et al., Aurora b is dispensable for megakaryocyte polyploidization, but contributes to the endomitotic process, Blood, 2010, vol. 116, no. 13, pp. 2345–2355.
Platica, M., Ionescu, A., Ivan, E., et al., Par, a protein involved in the cell cycle, is functionally related to chromosomal passenger proteins, Int. J. Oncol., 2011, vol. 38, no. 3, pp. 777–785.
Ratnam, S. Booth, R.L., et al., Endothelial cells from human and mice with polycystic kidney disease are characterized by polyploidy and chromosome segregation defects through surviving down-regulation, Hum. Mol. Genet., 2011, vol. 20, no. 2, pp. 354–367.
Takeuchi, M., Takeuchi, K., Ozawa, Y., et al., Aneuploidy in immortalized human mesenchymal stem cells with non-random loss of chromosome 13 in culture, In Vitro Cell Dev. Biol. Anim., 2009, vol. 45, nos. 5/6, pp. 290–299.
Illidge, T.M., Cragg, M.S., Fringes, B., et al., Polyploid giant cells provide a survival mechanism for p53 mutant cells after DNA damage, Cell Biol. Int., 2000, vol. 24, no. 9, pp. 621–633.
Walen, K.H., Bipolar genome reductional division of human near-senescent, polyploidy fibroblast cells, Cancer Genet. Cytogenet., 2007, vol. 173, no. 1, pp. 43–50.
Puig, P.E., Guilly, M.N., Bouchout, A., et al., Tumor cells can escape DNA-damaging cisplatin through DNA endoreduplication and reversible polyploidy, Cell Biol. Int., 2008, vol. 32, no. 9, pp. 1031–1010.
Ianzini, F., Kosmacek, E.A., Nelson, E.S., et al., Activation of meiosis-specific genes is associated with depolyploidication of human tumor cells following radiation-induced mitotic catastrophe, Cancer Res., 2009, vol. 69, no. 6, pp. 2296–2304.
Salmina, K., Jankevics, E., Huna, A., et al., Up-regulation of the embryonic self-renewal network through reversible polyploidy in irradiated p53-mutant tumour cells, Exp. Cell Res., 2010, vol. 316, no. 13, pp. 2099–2112.
Iatsyshyna, A.P., Glazko, T.T., Kovaleva, O.A., et al., The possible ways of the diploidization of the polyploid germinative stem cells in the mouse BALB/c line, Tsitol. Genet., 2006, vol. 40, no. 6, pp. 44–49.
Lukash, L.L., Mutagenesis in the integration processes and the evolution of the nuclear genome, Biopolym. Cell, 2007, vol. 23, no. 3, pp. 172–187.
King, R.W., When 2 + 2 = 5: the origins and fates of aneuploid and tetraploid cells, Biochim. Biophys. Acta, 2008, vol. 1786, no. 1, pp. 4–14.
Mosieniak, G. and Sikora, E., Polyploidy: the link between senescence and cancer, Curr. Pharm. Des., 2010, vol. 16, no. 6, pp. 734–740.
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Original Ukrainian Text © H.R. Akopian, N.L. Huleyuk, V.O. Kushniruk, D.O. Mykytenko, A.P. Yatsyshyna, L.L. Lukash, 2013, published in Tsitologiya i Genetika, 2013, Vol. 47, No. 5, pp. 55–69.
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Akopyan, H.R., Huleyuk, N.L., Kushniruk, V.O. et al. Comparative analysis of the karyotype of new human cell line 4BL at long-term cultivation: Ploidy of the chromosomal set. Cytol. Genet. 47, 305–317 (2013). https://doi.org/10.3103/S0095452713050022
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DOI: https://doi.org/10.3103/S0095452713050022


